67 results about "Cell factory" patented technology

The invention discloses a bacillus constitutive strong promoter and application thereof, and relates to the technical field of biology, and the nucleotide sequence of the promoter is SEQ ID NO: 2. The mRNA transcriptional level of the reporter gene is improved; the method is used for starting a target gene to efficiently express amylase in bacillus subtilis. The method is also used for starting a target gene to efficiently express alkaline protease in bacillus licheniformis. The novel constitutive promoter with enhanced transcriptional activity is developed by taking P43 as a starting template, and under the same host and culture conditions, the absolute level of the constitutive promoter for driving expression of a reporter gene or a target product is higher than that of the current P43 promoter, so that core power is provided for performance improvement of a bacillus cell factory.

The invention provides a cell strain for producing engineered viroid particles, a method thereof and the engineered viroid particles, the cell strain is named as SZCV-Con28 and preserved in Guangdong Microbial Culture Collection Center on October 15, 2025, the preservation number is GDMCC No: 67101, and the preservation number is GDMCC No: 67101. A plurality of corresponding cell factories for producing the engineered viroid particles are established by cell lines obtained by introducing different target gRNA elements into the cell strain, so that the corresponding engineered viroid particles can be continuously produced in batches. According to the invention, large-scale preparation can be realized through cell expanding culture, the capacity limitation is overcome, the production process flow is simplified, and the consumption of production raw materials is reduced.

The invention discloses curcuma wenyujin sesquiterpene synthase CwTPS8 as well as a coding gene and application thereof, and belongs to the technical field of plant genetic engineering, the amino acid sequence of the curcuma wenyujin sesquiterpene synthase CwTPS8 is shown as SEQ ID NO.1, the nucleotide sequence of the gene for coding the curcuma wenyujin sesquiterpene synthase CwTPS8 is shown as SEQ ID NO.2, and the nucleotide sequence of the gene for coding the curcuma wenyujin sesquiterpene synthase CwTPS8 is shown as SEQ ID NO.1. The curcuma wenyujin sesquiterpene synthase CwTPS8 can be used for preparing beta-caryophyllene, alpha-humulene and other curcuma wenyujin sesquiterpenoids by taking farnesyl pyrophosphate FPP as a substrate, and the technology can be used for subsequently producing the sesquiterpenoids through a microbial cell factory. Meanwhile, a foundation is laid for heterologous synthesis and further development and utilization of sesquiterpenoids such as beta-caryophyllene and alpha-humulene.

This invention aims to create a spatially isolated local reaction microenvironment for Rubisco by introducing a complete β-carboxysome microcompartment system. This design goal avoids oxygen inhibition, laying the foundation for ultimately achieving efficient and specific CO2 fixation. It provides a novel approach to overcome the fundamental problem of low efficiency of carbon-fixing enzymes in cytosols due to intense oxygen competition. By introducing the complete carboxysome system as a module, engineered yeast strains with high carboxylase activity and significantly improved ethanol production were obtained, providing a solid foundation for building next-generation yeast cell factories that produce high-yield ethanol using CO2 as a raw material.

The application discloses a kind of biological immobilized enzyme based on yeast surface display technology and the application of synthesis 2-phenylethanol, using yeast surface display technology, for the first time with Saccharomyces cerevisiae surface protein a-lectin as anchor protein, the key enzyme (transaminase, decarboxylase, ethanol dehydrogenase) involved in 2-PE producing Ehrlich pathway in Saccharomyces cerevisiae is separately or jointly carried out biological fixation, enhance substrate channel effect and proximity effect, in vitro one-step catalysis L-phenylalanine synthesis 2-PE, provide yield.The method for fixing multiple enzymes of Ehrlich pathway in the application can be used to reconfigure the biosynthesis pathway of other natural products in vitro, to provide feasible scheme for the rational design of cell factory.

The application belongs to the technical field of microbial sewage treatment, and particularly relates to an iron-carbon quantum dot / electrically active non-oxygen-producing photosynthetic bacteria / carrier hybrid cell factory and application thereof in sewage resourceization or carbon emission reduction; the iron-carbon quantum dot in the hybrid cell factory provided by the application promotes the non-oxygen-producing purple photosynthetic bacteria to utilize more light energy, and generates photo-generated electrons as an extracellular efficient electron donor source, and the photosensitive active substances in the extracellular polymeric substance of the non-oxygen-producing purple photosynthetic bacteria can promote the formation of a continuous cycle of trivalent iron and divalent iron, realize mutual cooperation, promote sewage resourceization and carbon emission reduction, and synthesize high-value-added products; and in addition to facilitating the recovery of biomass, the transparent conductive glass electrode as the carrier also ensures that the non-oxygen-producing purple photosynthetic bacteria stably and efficiently utilize light energy; therefore, the hybrid cell factory provided by the application is a high-efficiency microbial sewage treatment technology, and solves the technical problem of lack of high-efficiency microbial sewage treatment technology in the prior art.

The present disclosure relates to genetically modified host cells producing prostaglandins, to genetic constructs for expression of such mutants; to cultures of the genetically modified host cells and its use to produce the said products; to fermentation liquids comprising the said products resulting from such production; to compositions comprising the fermentation liquid; to pharmaceutical preparations made from such compositions and to the uses of such compositions and preparations.

The utility model relates to cell culture technical field discloses a kind of cell factory device, including plug body and the cover disc and culture tray of stacked arrangement, cover disc is located the top of culture tray, culture tray includes culture bottom plate, culture side plate and communicating part, culture side plate is arranged along the edge of culture bottom plate, and one end of culture side plate is connected with culture bottom plate and is enclosed to form culture cavity, and the other end of culture side plate is connected with culture tray, and culture tray seals culture cavity, culture bottom plate is equipped with through-hole, and communicating part is located in culture cavity, and one end of communicating part is connected in through-hole, and the other end of communicating part is formed with flow port between culture tray, cover disc is equipped with liquid injection hole, and the position of liquid injection hole is correspondingly set with the position of communicating part, and liquid injection hole is communicated with culture cavity by flow port, and plug body is sealedly connected with one end of communicating part away from cover disc.The cell factory device of the utility model improves the control of flatness of cell factory device, and ensures that cell growth is evenly distributed.

The application relates to a polymeric glutamic acid sugar ester and a biological fermentation preparation method and application thereof, and belongs to the technical field of biological fermentation. According to the application, amino acids and functional oligosaccharides are used as substrates, and a 'one-body cell factory' biosynthesis technology is designed to realize efficient directional assembly of high-molecular polymeric glutamic acid sugar esters. The application provides a brand-new polymeric glutamic acid sugar ester compound, and provides a biological fermentation method of the polymeric glutamic acid sugar ester, which is suitable for industrialized continuous production, and the prepared polymeric glutamic acid sugar ester has a narrower molecular weight distribution and better product uniformity.

The embodiment of the invention provides a cell factory reactor and a use method, and relates to the technical field of cell culture, the cell factory reactor comprises: a cell factory main body having a plurality of culture layers and a first channel respectively communicated with the plurality of culture layers, each culture layer having an upper surface and a lower surface, both the upper surface and the lower surface being subjected to TC treatment; the sensing assembly is used for acquiring culture environment parameters in the cell factory main body; the process gas supply assembly is connected with the first channel; the pipeline system comprises a liquid input assembly and a liquid output assembly, the liquid input assembly is used for conveying materials required by cell culture to the first channel, and the liquid output assembly is used for transferring liquid or cell suspension in the cell factory main body to the outside of the cell factory main body; the controller comprises a swinging mechanism, and the swinging mechanism is used for driving the cell factory main body to swing. According to the embodiment of the invention, the scale of cell amplification is increased, and the problem of insufficient amplification scale of a traditional cell factory is effectively solved.

The invention relates to the technical field of microbial fermentation and genetic engineering, and particularly discloses a yeast engineering bacterium for efficiently producing plastic depolymerases and application of the yeast engineering bacterium. The engineering yeast takes pichia pastoris as an expression host, a molecular chaperone is co-expressed, a vacuole sorting receptor gene is knocked out or knocked down, in addition, a yeast cell factory for efficiently expressing the plastic depolymerases is constructed by further combining regulation and control element optimization and target gene copy number regulation, and the engineering yeast has the advantages that the engineering yeast can be used for efficiently expressing the plastic depolymerases; the regulatory element comprises any one or a combination of more of a strong promoter, a signal peptide and a C-terminal tag. The engineering yeast provided by the invention can realize efficient accumulation and secretion of the plastic depolymerases, obviously improve the enzyme yield and reduce the production cost of unit enzyme activity, and provides a stable and efficient yeast expression system for industrial and low-cost production of the plastic depolymerases; the method has an important application prospect in the fields of biodegradation and resource utilization of waste plastics.

This invention relates to a Bacillus licheniformis strain with high tolerance to methanol and formaldehyde, belonging to the fields of synthetic biology and microbial engineering. This invention utilizes systematic mutagenesis and directed evolution to screen for a Bacillus licheniformis strain exhibiting excellent dual tolerance to methanol and formaldehyde. Bacillus licheniformis GW-MF, accession number CGMCC No. 38523. This strain can tolerate methanol concentrations of 80 g / L or even higher and formaldehyde concentrations of 0.02 g / L. This strain provides a key platform for constructing methanol-feeding cell factories, suitable for the green manufacturing process of methanol bioconversion to synthesize high-value-added products. It meets the development needs of a one-carbon bioeconomy under the "dual carbon" goal and has great application potential.

The application discloses a novel plasmid vector with improved CHO exogenous protein expression amount and construction and application thereof, and belongs to the technical field of genetic engineering. The plasmid vector pCHO6GS belongs to a novel plasmid system, and can at least improve the expression amount of general monoclonal antibodies and difficult-to-express recombinant proteins (such as H5HA trimer) in CHO by more than 1 times, that is, the unexpected improvement of the exogenous protein expression amount is realized by the novel plasmid vector, and the novel plasmid vector has an extremely important role in cost reduction and efficiency increase of various protein products with CHO as a cell factory.

This invention discloses a method for constructing a high-succinic acid-producing Pichia pastoris cell factory, using Pichia pastoris (… Komagataella phaffii Using GS115 as the chassis, a reductive TCA cycle enhancement strategy and an sdh5 knockout strategy were first constructed, respectively. It was found that a single strategy was insufficient to effectively achieve succinic acid accumulation. Further, the two strategies were combined to obtain the engineered strain SAOG, which achieved succinic acid accumulation. Based on this, the malate transporter SpMAE was introduced to enhance the transmembrane transport capacity of succinic acid, and histidine auxotrophic recovery was combined to improve the growth and fermentation performance of the strain, resulting in the engineered strain SH, whose succinic acid accumulation was 25.1% higher than that of SAOG. Finally, the engineered strain SH achieved a succinic acid yield of 10.47 g / L in shake-flask fermentation and 112.6 g / L after 120 h of continuous fed-batch fermentation in a 5 L fermenter. This invention achieves efficient succinic acid synthesis in the Pichia pastoris chassis through the synergistic regulation of reductive TCA pathway enhancement, succinic acid oxidation blocking, enhanced transmembrane transport, and auxotrophic recovery, exhibiting stable fermentation performance, high yield, and good industrial application potential.

The invention provides a biosynthesis method of 6-hydroxy-L-tryptophan, on the basis that a novel tryptophan 6-site hydroxylase protein family enzyme is obtained through screening, a cell factory is constructed for biotransformation, and green synthesis of 6-hydroxy-L-tryptophan is achieved. The technical problems of limited types of biocatalysts, insufficient chemical synthesis region selectivity and the like in the prior art are effectively solved. The amino acid sequence of the tryptophan 6-site hydroxylase is shown as SEQ ID NO: 1. The invention provides a protein sequence for catalyzing a new family of tryptophan 6-site hydroxylase, and 6-hydroxy-L-tryptophan can be directly and efficiently synthesized through a cell factory. The method has the advantages of mild reaction conditions, environmental friendliness, high regioselectivity, good product purity, flexible process route and the like, and a brand new technical scheme is provided for industrial production of 6-hydroxy-L-tryptophan.

The application discloses high-temperature-resistant engineered methanotrophs with high glutamic acid yield, a construction method and application thereof, and belongs to the field of industrial biotechnology. Methylotuvimicrobium buryatense 5GB1 as the host strain, transcriptional inhibition of the SOS response system by LexA protein is removed by knocking out a lexA gene in the genome of the host strain, expression of DNA damage repair related genes in the cell is activated, and high-temperature-resistant engineered methanotrophs MBΔ lexA that can stably grow in a high-temperature environment of 40-45 DEG C are obtained. The strain can still maintain normal growth and metabolism with methane as the sole carbon source under the condition of 40-45 DEG C, and the glutamic acid synthesis capacity of the strain is significantly better than that of the original strain at normal temperature. The high-temperature-resistant engineered methanotrophs MBΔ lexA provide important strain resources for the production of amino acid products by methanotroph cell factories in a high-temperature industrial scene, and are expected to improve the economy and sustainability of methanotrophic bioconversion.

The invention discloses a method for enhancing the synthesis capacity of a yeast double-metabolic-pathway pigment product and application, and belongs to the field of synthetic biology and metabolic engineering. The method comprises the following steps: constructing a saccharomyces cerevisiae engineering strain capable of simultaneously synthesizing violacein and lycopene; constructing a mutant library of the global transcription factor SPT15; transforming the mutant library into the engineering strain, and screening the mutant strain with increased yield based on bacterial colony color visualization. According to the invention, a single global transcription factor SPT15 is modified, so that the cell metabolism is regulated and controlled on the whole, the synergistic interaction of a diheterologous synthesis pathway is realized, and the tedious multi-gene rational modification is avoided. The method is easy and convenient to operate and short in period, and a new strategy is provided for efficient construction of yeast cell factories for high-yield pigment and other double-way products.

The present application relates to the field of microorganisms and genetic engineering, and specifically discloses an Escherichia coli engineering bacterium for efficiently producing plastic depolymerase and application thereof. The present application uses Escherichia coli as a host, and through synergistic optimization of multiple key dimensions such as signal peptide, promoter, solubility-promoting tag and co-expression of molecular chaperones, a multi-dimensional engineering system for efficiently expressing plastic depolymerase is constructed. The construction method provided by the present application includes a rational design and a screening strategy for combination and assembly of the multiple elements. The engineering bacterium can realize extracellular high-level expression of plastic depolymerase during induced fermentation, and the enzyme preparation prepared can be directly used for catalyzing efficient depolymerization of polyester plastics. The present application provides an Escherichia coli cell factory with stable high yield for industrialization and low-cost production of plastic depolymerase, and has important industrial application prospects for promoting large-scale biological degradation and resource recycling of waste plastics.

The application discloses an acetyl lactic acid synthase mutant and application thereof, and belongs to the technical field of bioactive enzymes. The acetyl lactic acid synthase mutant is acetyl lactic acid synthase-PCC6803-L255I, acetyl lactic acid synthase-PCC6803-V398I or acetyl lactic acid synthase-PCC6803-T488V. The application of the acetyl lactic acid synthase mutant in catalyzing preparation of acetyl lactic acid. The acetyl lactic acid synthase derived from PCC6803 is reformed by a site-directed mutation method, and mutant enzymes, acetyl lactic acid synthase-PCC6803-L255I, acetyl lactic acid synthase-PCC6803-V398I and acetyl lactic acid synthase-PCC6803-T488V, are screened, and the catalytic efficiency of the mutant enzymes is 120%, 130% and 110% of that of the wild enzyme respectively. The application lays a technical foundation for metabolic engineering of a blue-green algae photosynthetic cell factory.

This invention discloses a directed evolution strategy for SsDMS, a type II sesquiterpene cyclase derived from *Streptomyces showdoensis*, based on alanine scanning, and its applications. The invention describes several key mutant sites in SsDMS capable of catalyzing the generation of novel dried sesquiterpene compounds, including substitutions at positions 208, 248, 249, 497, and 505. This invention also discloses a highly efficient *E. coli* cell factory applied to the heterologous expression of SsDMS mutants and the efficient production of novel dried sesquiterpene compounds, further yielding structurally novel dried sesquiterpene compounds. This invention achieves the efficient production of dried sesquiterpene compounds using SsDMS and its mutants. Its modification strategy has broad applicability, providing important enzymatic resources for the biosynthesis of novel terpenoids and offering more options for the discovery of drug lead compounds.

The present application relates to a kind of methyltransferase that can catalyze anthraquinone compound methylation, amino acid sequence is as shown in SEQ ID NO:2, also provide the application of this methyltransferase in the production of emodin-3-methyl ether and other methylated anthraquinone compounds.The present application digs a new methyltransferase, which can methylate anthraquinone compound, and can convert emodin into emodin-3-methyl ether.The present application constructs the engineering bacteria that can produce emodin-3-methyl ether using the coding gene of the enzyme, widens the methyltransferase library that can be used for emodin methylation, provides new way for emodin-3-methyl ether cell factory construction.

The utility model discloses automatic production equipment for cell drugs. The automatic production equipment comprises a process operation cavity and a cell factory incubator, the cell factory incubator is communicated with the process operation cavity; the process operation cavity is provided with a cell harvesting area and a cell inoculation liquid changing area; a centrifugal bottle pipetting and cover opening and closing assembly, a centrifugal assembly, a subpackaging assembly and a first transfer mechanism are arranged in the cell harvesting area; the first transfer mechanism is used for transferring the centrifugal bottle between the centrifugal bottle pipetting and cover opening and closing assembly and the centrifugal assembly; a cell factory pipetting and cover opening and closing assembly and a second transfer mechanism are arranged in the cell inoculation liquid changing area; the second transfer mechanism is used for transferring the cell factory among the cell factory pipetting and cover opening and closing assembly, the cell factory incubator and the centrifugal bottle pipetting and cover opening and closing assembly. By means of the arrangement, the function of full-process automation is achieved, and the purposes of reducing manual intervention and improving operation efficiency and sterility are achieved.

The invention discloses a valene synthase mutant and application thereof in biosynthesis of valene, and belongs to the field of enzyme engineering and synthetic biology. The invention utilizes synthetic biology and enzyme engineering means to obtain the varene synthase mutant and the application of the varene synthase mutant in synthesis of varene in yeast, and by providing the varene synthase mutant with improved catalytic activity, the yield of varene is improved, and conditions are created for industrial production of varene. Compared with the currently reported high-activity valarene synthase EgVSm, the valarene synthase mutant EgVSmK400H-H487G-F50A-V185F disclosed by the invention has higher activity, the yield of the valarene is improved by 50%, the yield level of biosynthesized valarene is further improved, and a foundation is laid for constructing a yeast high-yield valarene cell factory.

The invention discloses recombinant escherichia coli with high yield of cyanidin-3-O-glucoside as well as a construction method and application of the recombinant escherichia coli. The method comprises the following steps: by taking escherichia coli BL21 (DE3) engineering bacteria as a host, integrating flavanonol 4-reductase, anthocyanin synthetase and flavonoid 3-O-glucosyltransferase from different sources onto a plasmid vector pRSFDuet, and constructing to obtain a chassis cell for producing C3G; meanwhile, glutathione transferase is overexpressed, dissolution-promoting tag small ubiquitin-like modified protein is added, FaDFR is modified through enzyme engineering, FaDFR catalytic efficiency is improved through artificial protein cage space combination pathway enzyme, key precursor UDP-glucose is synthesized in an in-situ intensified mode and cofactor engineering, the cell factory for efficiently synthesizing C3G is constructed based on the combination strategy, the yield of C3G is increased, and the method has the advantages that the method is simple and convenient to operate, and the cost is low. The titer reaches 797.20 mg / L. The method provided by the invention has the characteristics of simplicity, economy and eco-friendliness.

The invention discloses a cell factory production method and application of an IBDV recombinant MDV vector live vaccine, and belongs to the technical field of veterinary vaccine preparation. In order to overcome the defects existing in traditional recombinant MDV vaccine production and solve the technical problems that a cell factory production process of an IBDV recombinant MDV vector live vaccine lacks system optimization and the like, the method comprises the steps of process parameter optimization, virus titer detection, immune effect verification and genetic stability evaluation. The invention provides a process method for preparing an IBDV recombinant MDV vector live vaccine by using a cell factory system, and the virus titer and hereditary stability are remarkably improved, so that the production efficiency and immune effect of the vaccine are improved.

The application discloses a cembranoid alcohol synthase mutant and application thereof in biosynthesis of cembranoid alcohol, and belongs to the field of enzyme engineering and synthetic biology. The mutant with significantly improved catalytic performance is obtained by rational design and mutation screening of cembranoid alcohol synthase (NtCBTS) from tobacco. The cembranoid alcohol yield of the strain containing the mutant NtCBTS_K208R-A268C-K353R is increased by 231.18% compared with the strain containing NtCBTS. The mutant is applied to a Saccharomyces cerevisiae expression system, and efficient biosynthesis of cembranoid alcohol is realized, and the generation level of the target product is significantly improved. The high-activity enzyme mutant and the application strategy provided by the application effectively improve the biosynthesis efficiency of cembranoid alcohol, and provide important technical support for constructing a high-yield yeast cell factory and promoting industrialized production.

Disclosed are a novel nuclear localization sequence (NLS) mutant and a method for improving biosynthetic efficiency using same, belonging to the field of bioengineering. According to the present disclosure, a novel NLS is screened out, and recombinant yeast with improved metabolite synthesis efficiency is constructed by applying the NLS obtained through screening, which provides key technical support for developing a high-yield, stable and cost-effective yeast biosynthesis platform, and facilitates the use of recombinant yeast as a cell factory for large-scale industrial biomanufacturing.

The invention discloses an optimization method and application of sclareol production strains based on various metabolic models. The method comprises the following steps: integrating phenotype data and transcriptome data of a chassis strain to construct an integration model, and predicting an endogenous transformation target within a genome range based on a metabolism model; the host strain is selected from saccharomyces cerevisiae. According to the method, an enzyme constraint model and a non-intuitive gene editing target mining strategy are adopted, and a new method and a new modified target are provided for constructing a microbial cell factory for efficiently synthesizing sclareol.

The invention provides a biosynthesis method of 7-hydroxy-L-tryptophan and a derivative thereof. The amino acid sequence of tryptophan 7-site hydroxylase used in the method is SEQ ID NO: 1. The invention provides an enzyme for catalyzing 7-site hydroxylation of L-tryptophan and a derivative thereof. The 7-hydroxy-L-tryptophan and the derivative thereof can be directly and efficiently synthesized through a cell factory. The synthesis mode of the 7-hydroxy-L-tryptophan and the derivative thereof has the characteristics of greenness and high efficiency, and provides an important solution strategy for site selective catalysis of 7-site hydroxylation of L-tryptophan and the derivative thereof.

The utility model relates to the technical field of cell culture in biopharmaceutical production, in particular to a novel 40-layer cell factory with a metal net interlayer, which is characterized in that the metal net interlayer is designed in a partition plate between culture layers of the original 40-layer cell factory and comprises a metal net structure and an internal temperature sensor component; and a T-shaped all-metal sheet covered edge is arranged on the peripheral edge of the layer of the metal net-shaped structure. According to the novel design structure, uniform temperature rise of each cell culture layer can be realized, the temperature rise efficiency is high, temperature values at different positions can be conveniently monitored, and the stability and controllability of biological products and vaccine products taking a 40-layer cell factory as a cell or virus culture container in the research and development, pilot plant test and industrial production processes are greatly improved.